CN110016493A - It is a kind of cause high sugar product swell microorganism separation and bacteriostasis method - Google Patents
It is a kind of cause high sugar product swell microorganism separation and bacteriostasis method Download PDFInfo
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- CN110016493A CN110016493A CN201910356010.1A CN201910356010A CN110016493A CN 110016493 A CN110016493 A CN 110016493A CN 201910356010 A CN201910356010 A CN 201910356010A CN 110016493 A CN110016493 A CN 110016493A
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- 244000005700 microbiome Species 0.000 title claims abstract description 104
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000000926 separation method Methods 0.000 title claims abstract description 18
- 239000001963 growth medium Substances 0.000 claims abstract description 101
- 238000009629 microbiological culture Methods 0.000 claims abstract description 54
- 230000006872 improvement Effects 0.000 claims abstract description 40
- 239000003755 preservative agent Substances 0.000 claims abstract description 40
- 230000002335 preservative effect Effects 0.000 claims abstract description 33
- 241000233866 Fungi Species 0.000 claims abstract description 20
- 238000012797 qualification Methods 0.000 claims abstract description 15
- 239000002609 medium Substances 0.000 claims description 22
- 239000012895 dilution Substances 0.000 claims description 20
- 238000010790 dilution Methods 0.000 claims description 20
- WXMKPNITSTVMEF-UHFFFAOYSA-M sodium benzoate Chemical compound [Na+].[O-]C(=O)C1=CC=CC=C1 WXMKPNITSTVMEF-UHFFFAOYSA-M 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 16
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical class N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 15
- 235000010234 sodium benzoate Nutrition 0.000 claims description 15
- 239000004299 sodium benzoate Substances 0.000 claims description 15
- 230000001954 sterilising effect Effects 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 12
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- 239000001967 plate count agar Substances 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 7
- NVNLLIYOARQCIX-MSHCCFNRSA-N Nisin Chemical compound N1C(=O)[C@@H](CC(C)C)NC(=O)C(=C)NC(=O)[C@@H]([C@H](C)CC)NC(=O)[C@@H](NC(=O)C(=C/C)/NC(=O)[C@H](N)[C@H](C)CC)CSC[C@@H]1C(=O)N[C@@H]1C(=O)N2CCC[C@@H]2C(=O)NCC(=O)N[C@@H](C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(NCC(=O)N[C@H](C)C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCSC)C(=O)NCC(=O)N[C@H](CS[C@@H]2C)C(=O)N[C@H](CC(N)=O)C(=O)N[C@H](CCSC)C(=O)N[C@H](CCCCN)C(=O)N[C@@H]2C(N[C@H](C)C(=O)N[C@@H]3C(=O)N[C@@H](C(N[C@H](CC=4NC=NC=4)C(=O)N[C@H](CS[C@@H]3C)C(=O)N[C@H](CO)C(=O)N[C@H]([C@H](C)CC)C(=O)N[C@H](CC=3NC=NC=3)C(=O)N[C@H](C(C)C)C(=O)NC(=C)C(=O)N[C@H](CCCCN)C(O)=O)=O)CS[C@@H]2C)=O)=O)CS[C@@H]1C NVNLLIYOARQCIX-MSHCCFNRSA-N 0.000 claims description 6
- 108010053775 Nisin Proteins 0.000 claims description 6
- 235000010297 nisin Nutrition 0.000 claims description 6
- 239000004309 nisin Substances 0.000 claims description 6
- CHHHXKFHOYLYRE-UHFFFAOYSA-M 2,4-Hexadienoic acid, potassium salt (1:1), (2E,4E)- Chemical compound [K+].CC=CC=CC([O-])=O CHHHXKFHOYLYRE-UHFFFAOYSA-M 0.000 claims description 5
- 235000021433 fructose syrup Nutrition 0.000 claims description 5
- 235000010241 potassium sorbate Nutrition 0.000 claims description 5
- 239000004302 potassium sorbate Substances 0.000 claims description 5
- 229940069338 potassium sorbate Drugs 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 239000000654 additive Substances 0.000 claims description 4
- 230000000996 additive effect Effects 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 229930187593 rose bengal Natural products 0.000 claims description 3
- AZJPTIGZZTZIDR-UHFFFAOYSA-L rose bengal Chemical compound [K+].[K+].[O-]C(=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1C1=C2C=C(I)C(=O)C(I)=C2OC2=C(I)C([O-])=C(I)C=C21 AZJPTIGZZTZIDR-UHFFFAOYSA-L 0.000 claims description 3
- 229940081623 rose bengal Drugs 0.000 claims description 3
- STRXNPAVPKGJQR-UHFFFAOYSA-N rose bengal A Natural products O1C(=O)C(C(=CC=C2Cl)Cl)=C2C21C1=CC(I)=C(O)C(I)=C1OC1=C(I)C(O)=C(I)C=C21 STRXNPAVPKGJQR-UHFFFAOYSA-N 0.000 claims description 3
- 238000013461 design Methods 0.000 abstract description 4
- 235000019249 food preservative Nutrition 0.000 abstract description 4
- 239000005452 food preservative Substances 0.000 abstract description 4
- 239000000047 product Substances 0.000 description 42
- 235000020434 chocolate syrup Nutrition 0.000 description 8
- 239000013078 crystal Substances 0.000 description 8
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 235000013361 beverage Nutrition 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 5
- 230000001408 fungistatic effect Effects 0.000 description 5
- 230000002401 inhibitory effect Effects 0.000 description 5
- 238000012216 screening Methods 0.000 description 5
- 239000002253 acid Substances 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 239000012898 sample dilution Substances 0.000 description 4
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000019634 flavors Nutrition 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 229930091371 Fructose Natural products 0.000 description 2
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 2
- 239000005715 Fructose Substances 0.000 description 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 2
- 239000008373 coffee flavor Substances 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000007863 gel particle Substances 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 229910052708 sodium Inorganic materials 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 238000012795 verification Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 239000013065 commercial product Substances 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 230000004992 fission Effects 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 235000010235 potassium benzoate Nutrition 0.000 description 1
- 239000004300 potassium benzoate Substances 0.000 description 1
- 229940103091 potassium benzoate Drugs 0.000 description 1
- CHHHXKFHOYLYRE-STWYSWDKSA-M potassium sorbate Chemical compound [K+].C\C=C\C=C\C([O-])=O CHHHXKFHOYLYRE-STWYSWDKSA-M 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 238000012859 sterile filling Methods 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000012976 tarts Nutrition 0.000 description 1
- 238000012549 training Methods 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
- C12Q1/045—Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Toxicology (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of microorganism separation for causing high sugar product swell and bacteriostasis methods, the pol of microbiological culture media are adjusted to high pol 20Brix~60Brix including (1), pH is adjusted to 4.0~6.0, obtains the microbiological culture media of improvement；(2) inverted plate or plate streaking are mixed, microculture is carried out；(3) single colonie for selecting different shape carries out microorganism fungus kind identification, obtains qualification result, finds the suspicious microorganism for causing swell, access in the non-swell sample of high sugar product and cultivated, and sees whether meeting swell, if so, being swell microorganism；(4) different preservatives being separately added into the microbiological culture media of the fresh improvement at more parts and being inoculated with swell microorganism, cultivate and observed whether there is or not growth, selection inhibits the preservative of its growth.The present invention can quickly screen the objective microbe for causing swell, and obtain effective food preservative, ingenious in design to prevent the generation of swell, simple to operate, at low cost.
Description
Technical field
The present invention relates to food technology field, in particular to high sugar product technical field, in particular to one kind cause high sugar
The microorganism of product swell separates and bacteriostasis method.
Background technique
High candy juice refers in product based on sugar, inspissated juice, Normal juice, adds or do not add food sweetener, tart flavour
Agent, pigment, essence, product made of preservative.The factory of the long-term production product has a liking for the microbes of high sugar by selectivity
Existence in the environment, microorganism is ubiquitous in the environment, if not sterile filling production line, is easy to cause this hobby
High sugared or resistance to hypertonic microorganism remaining is in the product or secondary pollution product, the swell being visually observed belong to the micro- life of swell
Object largely breeds the phenomenon that causing.So how to utilize existing product line, product yield is improved to greatest extent, is reduced public
Department's loss is a urgent problem.
Therefore, it on the basis of apparatus for production line and technique cannot be modified, to prevent the generation of product swell, need to find
Cause the objective microbe of swell, and use effective food preservative, to prevent the generation of swell.
Summary of the invention
In order to overcome the disadvantages of the prior art mentioned above, it is an object of the present invention to provide one kind to cause high sugar product
The microorganism of swell separates and bacteriostasis method, can quickly screen the objective microbe for causing swell, and obtain effective food
Preservative is suitable for large-scale promotion application to prevent the generation of swell.
The microorganism separation for causing high sugar product swell another object of the present invention is to provide a kind of and bacteriostasis method,
It is ingenious in design, it is simple to operate, it is at low cost, it is suitable for large-scale promotion application.
To achieve the above objectives, the present invention provide it is a kind of cause high sugar product swell microorganism separation and bacteriostasis method,
Its main feature is that, comprising the following steps:
(1) pol of microbiological culture media is adjusted to high pol 20Brix~60Brix, by the microbiological culture media
PH adjust to 4.0~6.0, obtain the microbiological culture media of improvement；
(2) by the microbiological culture media mixing inverted plate of the swell sample of high sugar product and the improvement, or by institute
The swell sample for the high sugar product stated is crossed on the plate of the microbiological culture media of the improvement, and microculture is carried out；
(3) single colonie for selecting different shape carries out microorganism fungus kind identification, obtains microorganism fungus kind qualification result, according to
The microorganism fungus kind qualification result finds the suspicious microorganism for causing swell, by the suspicious microorganism access high sugar
Cultivated in the non-swell sample of product, observe the high sugar product non-swell sample whether can swell, if so, institute
Stating suspicious microorganism is swell microorganism；
(4) different preservatives is separately added into the microbiological culture media of the fresh improvement at more parts and be inoculated with institute
Swell microorganism is stated, cultivate and observes the swell microorganism whether there is or not growth, selection inhibits the institute of the swell microorganism growth
State preservative.
Preferably, the step (1) specifically includes the following steps:
(11) the solid component mixture of microbiological culture media described in 1 parts by weight is taken, 7 parts by weight~21 parts by weight are added
The HCl solution of water and 0 parts by weight~0.2 parts by weight 1mol/L, is uniformly dissolved, obtains liquid medium；
(12) 1 parts by weight fructose syrup is taken, 0.04 other parts by weight~0.4 parts by weight water is added, is uniformly dissolved, obtains
To liquid glucose；
(13) liquid medium and the liquid glucose are individually positioned in high-pressure sterilizing pot 115 DEG C~121 DEG C, 15min
~20min sterilizing cools the mass ratio of rear 1:15~34 and mixes the liquid medium and the liquid glucose.
Preferably, the microbiological culture media is plate count agar culture medium or rose-bengal in the step (1)
Culture medium.
Preferably, the step (2) specifically includes the following steps:
(21) 10 times of gradient dilutions of the swell sample of the high sugar product are obtained into dilution；
(22) microbiological culture media mixing inverted plate of the dilution described in 1ml with the improvement is taken, or by 100 microlitres
~200 microlitres of dilution scribing line are on the plate of the microbiological culture media of the improvement；
(23) 35 DEG C are cultivated 1 day~3 days.
Preferably, the pH of the microbiological culture media of the fresh improvement is 4 or 6 in the step (4).
Preferably, the additive amount of the preservative is 0.01 parts by weight~0.1 parts by weight in the step (4).
Preferably, the preservative is selected from poly- bad ammonia salt hydrochlorate, nisin, sorbic acid in the step (4)
It is two or more in potassium and sodium benzoate.
Beneficial effects of the present invention essentially consist in that:
1, the of the invention microorganism separation for causing high sugar product swell and bacteriostasis method include (1) by microbiological culture media
Pol be adjusted to high pol 20Brix~60Brix, pH is adjusted to 4.0~6.0, obtains the microbiological culture media of improvement；(2)
It is drawn by the microbiological culture media mixing inverted plate of the swell sample of high sugar product and improvement, or by the swell sample of high sugar product
Line carries out microculture on the plate of the microbiological culture media of improvement；(3) single colonie for selecting different shape carries out micro- life
Object strain idenfication obtains microorganism fungus kind qualification result and finds according to microorganism fungus kind qualification result and cause the suspicious micro- of swell
Suspicious microorganism is accessed in the non-swell sample of high sugar product and is cultivated by biology, observes the non-swell sample of high sugar product
Whether can swell, if so, suspicious microorganism be swell microorganism；(4) in the microbiological culture media of the fresh improvement at more parts
It is separately added into different preservatives and is inoculated with swell microorganism, cultivate and observe swell microorganism whether there is or not growth, selection inhibits swollen
Therefore the preservative of tank microorganism growth can quickly screen the objective microbe for causing swell, and it is anti-to obtain effective food
Rotten agent is suitable for large-scale promotion application to prevent the generation of swell.
2, the of the invention microorganism separation for causing high sugar product swell and bacteriostasis method include (1) by microbiological culture media
Pol be adjusted to high pol 20Brix~60Brix, pH is adjusted to 4.0~6.0, obtains the microbiological culture media of improvement；(2)
It is drawn by the microbiological culture media mixing inverted plate of the swell sample of high sugar product and improvement, or by the swell sample of high sugar product
Line carries out microculture on the plate of the microbiological culture media of improvement；(3) single colonie for selecting different shape carries out micro- life
Object strain idenfication obtains microorganism fungus kind qualification result and finds according to microorganism fungus kind qualification result and cause the suspicious micro- of swell
Suspicious microorganism is accessed in the non-swell sample of high sugar product and is cultivated by biology, observes the non-swell sample of high sugar product
Whether can swell, if so, suspicious microorganism be swell microorganism；(4) in the microbiological culture media of the fresh improvement at more parts
It is separately added into different preservatives and is inoculated with swell microorganism, cultivate and observe swell microorganism whether there is or not growth, selection inhibits swollen
The preservative of tank microorganism growth, it is therefore, ingenious in design, it is simple to operate, it is at low cost, it is suitable for large-scale promotion application.
These and other objects of the invention, feature and advantage, are filled by following detailed descriptions and claim
Fission is existing, and can be achieved by means, device and the their combination specially pointed out in appended claims.
Detailed description of the invention
Fig. 1 is using the microbiological culture media of improvement and the microbiological culture media that do not improve using mixing inverted plate method to swollen
The separating resulting of tank microorganism.
Fig. 2 is using the microbiological culture media of improvement and the microbiological culture media that do not improve using plate streak to swell
The separating resulting of microorganism.
Specific embodiment
For the generation for solving product swell, the present invention is by adjusting its Brix and pH on the basis of microbiological culture media
Whole, screening obtains the objective microbe for causing swell, and is chosen by different preservatives to the different inhibitory effects of objective microbe
Choosing obtains effective preservative.
Specifically, the present invention provides a kind of microorganism separation for causing high sugar product swell and bacteriostasis method, including following
Step:
(1) pol of microbiological culture media is adjusted to high pol 20Brix~60Brix, by the microbiological culture media
PH adjust to 4.0~6.0, obtain the microbiological culture media of improvement；
(2) by the microbiological culture media mixing inverted plate of the swell sample of the high sugar product and the improvement, or
The swell sample of the high sugar product is crossed on the plate of the microbiological culture media of the improvement, microorganism training is carried out
It supports；
(3) single colonie for selecting different shape carries out microorganism fungus kind identification, obtains microorganism fungus kind qualification result, according to
The microorganism fungus kind qualification result finds the suspicious microorganism for causing swell, by the suspicious microorganism access high sugar
Cultivated in the non-swell sample of product, observe the high sugar product non-swell sample whether can swell, if so, institute
Stating suspicious microorganism is swell microorganism；
(4) different preservatives is separately added into the microbiological culture media of the fresh improvement at more parts and be inoculated with institute
Swell microorganism is stated, cultivate and observes the swell microorganism whether there is or not growth, selection inhibits the institute of the swell microorganism growth
State preservative.
The step (1) can specifically include any suitable step, preferably, the step (1) specifically include it is following
Step:
(11) the solid component mixture of microbiological culture media described in 1 parts by weight is taken, 7 parts by weight~21 parts by weight are added
The HCl solution of water and 0 parts by weight~0.2 parts by weight 1mol/L, is uniformly dissolved, obtains liquid medium；
(12) 1 parts by weight fructose syrup is taken, 0.04 other parts by weight~0.4 parts by weight water is added, is uniformly dissolved, obtains
To liquid glucose；
(13) liquid medium and the liquid glucose are individually positioned in high-pressure sterilizing pot 115 DEG C~121 DEG C, 15min
~20min sterilizing cools the mass ratio of rear 1:15~34 and mixes the liquid medium and the liquid glucose.
In the step (1), the microbiological culture media can be any suitable microbiological culture media, preferably,
In the step (1), the microbiological culture media is plate count agar culture medium or rose bengal medium.
The step (2) can specifically include any suitable step, preferably, the step (2) specifically include it is following
Step:
(21) 10 times of gradient dilutions of the swell sample of the high sugar product are obtained into dilution；
(22) microbiological culture media mixing inverted plate of the dilution described in 1ml with the improvement is taken, or by 100 microlitres
~200 microlitres of dilution scribing line are on the plate of the microbiological culture media of the improvement；
(23) 35 DEG C are cultivated 1 day~3 days.
In the step (4), the pH of the microbiological culture media of the fresh improvement can according to need determination,
Preferably, the pH of the microbiological culture media of the fresh improvement is 4 or 6 in the step (4).
The additive amount of the preservative can according to need determination, preferably, in the step (4), the preservative
Additive amount be 0.01 parts by weight~0.1 parts by weight.
The preservative can be any suitable preservative, preferably, in the step (4), the preservative choosing
Autohemagglutination relies two or more in ammonia salt hydrochlorate, nisin, potassium sorbate and sodium benzoate.
In order to be more clearly understood that technology contents of the invention, spy lifts following embodiment and is described in detail.In the present invention,
Related component or raw material are conventional commercial product, or can be obtained by ordinary skill in the art means.
1 improved culture medium of embodiment screens swell microorganism and effective preservative screens
(1) prepare improvement solid medium: take 1 parts by weight PCA (plate count agar culture medium) (the rich biology in Qingdao sea,
250g/ bottles) culture medium, the water of 21 parts by weight is added, the HCl solution of 0.2 parts by weight 1mol/L is added, cone is added after being uniformly dissolved
Shape bottle, obtains liquid medium；1 parts by weight fructose syrup (F55 fructose, Jia Ji, tank car packaging, 30 tons/vehicle) is taken, 0.4 weight is added
Part water is measured, conical flask is added after being uniformly dissolved and obtains liquid glucose；Two kinds of liquid are individually positioned in high-pressure sterilizing pot and sterilize, and 121 DEG C,
15min, after sterilizing cools, 1:15 mixing, for use；Final culture medium Brix 22-25, pH 4.0-4.2.
(2) sample dilution and strain separating: by coffee crystal, (1.0kg/ containing gel particle bottles of coffee flavor beverage (fresh and alive
Fruit juice Co., Ltd)；Brix39-41 10 times of gradient dilutions of swell sample), after taking sample 1ml and the cooling of different dilutions
Culture medium mixing inverted plate, while taking plate made of the sample scribing line culture medium of 100 microlitres of different gradients；Plate is placed in 35
DEG C incubator culture 3 days.
(3) fungi preservation is with identification: different shape single colonie being separated, the identification of Song Waijian mechanism obtains microorganism fungus kind
Qualification result.According to microorganism fungus kind qualification result, the suspicious microorganism for causing swell is found, which is accessed into coffee
Cultivated in the non-swell sample of coffee crystal, observe coffee crystal non-swell sample whether can swell, if so, suspicious micro-
Biology is swell microorganism.
(4) effectively preservative screening: on the basis of improved culture medium, medium pH is adjusted respectively to 6.0,4.0；Brix
It adjusts to 22~25,0.01 parts by weight~0.1 parts by weight variety classes preservative is added, and be inoculated with swell microorganism, through 35
DEG C incubator culture 3 days, the fungistatic effect of preservative is observed, the results are shown in Table 1.
1 preservative of the table verification result at pH6.0 and pH4.0 respectively
| Preservative | pH4.0 | pH6.0 |
| The poly- bad ammonia salt hydrochlorate of 0.01 part of ε- | —/+/- | —/+/- |
| The poly- bad ammonia salt hydrochlorate of 0.02 part of ε- | —/+/- | —/+/- |
| The poly- bad ammonia salt hydrochlorate of 0.04 part of ε- | — | — |
| The poly- bad ammonia salt hydrochlorate of 0.06 part of ε- | — | — |
| The poly- bad ammonia salt hydrochlorate of 0.08 part of ε- | — | — |
| 0.02 part of nisin | ++ | ++ |
| 0.05 part of potassium sorbate | + | + |
| 0.1 part of sodium benzoate | — | + |
| Blank | ++ | ++ |
Note: "+" indicates obvious growth；" ++ " growth is clearly；"-" is without growth；" +/- " growth is unobvious.
The non-improved culture medium of comparative example 1 screens swell microorganism
(1) it prepares solid medium: taking 1 parts by weight PCA (plate count agar culture medium) (the rich biology in Qingdao sea, 250g/
Bottle) culture medium, the water of 21 parts by weight is added, the HCl solution of 0.2 parts by weight 1mol/L is added, conical flask is added after being uniformly dissolved,
Obtain liquid medium；It is placed on high-pressure sterilizing pot to sterilize, 121 DEG C, 15min, for use；Take 15 parts by weight sterile waters and 1 parts by weight
Above-mentioned culture medium mixing, final culture medium Brix2-3, pH 4.0-4.2.
(2) sample dilution and strain separating: by coffee crystal, (1.0kg/ containing gel particle bottles of coffee flavor beverage (fresh and alive
Fruit juice Co., Ltd) Brix 39-41) 10 times of gradient dilutions of swell sample, take different dilutions sample 1ml and it is cooling after
Culture medium mixing inverted plate, while taking plate made of the sample scribing line culture medium of 100 microlitres of different gradients；Plate is placed in 35
DEG C incubator culture 3 days.
Comparative example 1 the difference from embodiment 1 is that: do not adjust culture medium Brix to 22~25.
Carry out compliance test result to improvement and non-improved culture medium: referring to Figure 1 and shown in Fig. 2, improved culture medium can be a large amount of
It screens the microorganism for causing swell, rather than improved culture medium screening has no that microorganism grows, illustrates improvement to culture medium i.e.
Brix adjustment is the key point of swell microorganism separation.
Cause product swollen from Fig. 1-Fig. 2 and table 1 it is found that culture medium made from the embodiment of the present invention 1 can be clearly separated out
The microorganism of tank；And comparative example 1 has no that microorganism obviously grows.On the basis of improved culture medium, test under condition of different pH
Preservative effectiveness relies ammonia salt hydrochlorate to have apparent fungistatic effect it is found that pH 0.01 parts by weight under acid and neutrallty condition are poly-,
Sodium benzoate can play the effect for inhibiting swell microorganism in acid condition, and potassium sorbate and nisin are micro- to swell
It is biological invalid.
In order to examine the actual effect for inhibiting swell, by 2.0 × 105Cfu/ml swell microorganism is linked into pH4.0's
In coffee crystal, addition or do not add 0.1 part of sodium benzoate, 35 DEG C insulating box culture 1 month, the results are shown in Table 2, swell microorganism
100% causes the coffee crystal swell for not adding preservative sodium benzoate, and the coffee crystal for adding sodium benzoate has no swell.
Influence whether adding sodium benzoate in 2 coffee crystal of table to swell
| Project | Access strain viable count/cfu/ml | Swell quantity/ |
| 0.1 parts by weight sodium | 2.0×105 | 0/50 |
| Do not add sodium benzoate | 2.0×105 | 50/50 |
Embodiment 2 adjusts improved culture medium Brix to 60
(1) it prepares improvement fluid nutrient medium: taking 1 parts by weight PCA (plate count culture medium) (the rich biology in Qingdao sea, 250g/
Bottle) culture medium, the water of 7 parts by weight is added, the NaOH solution of 0.2 parts by weight 1mol/L is added, conical flask is added after being uniformly dissolved,
Obtain liquid medium；1 parts by weight fructose syrup (F55 fructose, Jia Ji, tank car packaging, 30 tons/vehicle) is taken, 0.04 parts by weight are added
Water is added conical flask and obtains liquid glucose after being uniformly dissolved；Two kinds of liquid are individually positioned in high-pressure sterilizing pot and sterilize, and 115 DEG C, 20min,
After sterilizing cools, 1:34 mixing, for use；Final culture medium Brix 60, pH6.0.
(2) sample dilution and strain separating: by chocolate syrup (Brix60) (chocolate syrup flavor beverage, 2L/ bottles
(fresh and alive fruit juice Co., Ltd), Brix 59-61) 10 times of gradient dilutions of swell sample, take the sample 1ml of different dilutions with
Culture medium mixing inverted plate after cooling, while taking plate made of the sample scribing line culture medium of 200 microlitres of different gradients；Plate
It is placed in 35 DEG C of incubator cultures 3 days.
(3) fungi preservation is with identification: different shape single colonie being separated, the identification of Song Waijian mechanism obtains microorganism fungus kind
Qualification result.According to microorganism fungus kind qualification result, the suspicious microorganism for causing swell is found, which is accessed skilful
Cultivated in the non-swell sample of gram force syrup, observe chocolate syrup non-swell sample whether can swell, if so, can
Doubting microorganism is swell microorganism.
(4) effectively preservative screening: on the basis of improved culture medium, above-mentioned medium pH is adjusted respectively to 6.0,4.0；
Brix is adjusted to 60,0.01 parts by weight~0.1 parts by weight variety classes preservative is added, and be inoculated with swell microorganism, through 35
DEG C incubator culture 3 days, the fungistatic effect of preservative is observed, the results are shown in Table 3.
3 preservative of the table verification result at pH6.0 and pH4.0 respectively
| Preservative | pH4.0 | pH6.0 |
| The poly- bad ammonia salt hydrochlorate of 0.01 part of ε- | — | — |
| The poly- bad ammonia salt hydrochlorate of 0.02 part of ε- | — | — |
| The poly- bad ammonia salt hydrochlorate of 0.04 part of ε- | — | — |
| The poly- bad ammonia salt hydrochlorate of 0.06 part of ε- | — | — |
| The poly- bad ammonia salt hydrochlorate of 0.08 part of ε- | — | — |
| 0.02 part of nisin | + | + |
| 0.05 part of potassium sorbate | — | + |
| 0.1 part of sodium benzoate | — | + |
| Blank | ++ | ++ |
Note: "+" indicates obvious growth；" ++ " growth is clearly；"-" is without growth.
The non-improved culture medium of comparative example 2 screens swell microorganism
(1) it prepares fluid nutrient medium: taking 1 parts by weight PCA (plate count culture medium) (the rich biology in Qingdao sea, 250g/ bottles),
The water of 7 parts by weight is added, the NaOH solution of 0.2 parts by weight 1mol/L is added, conical flask is added after being uniformly dissolved, obtains culture medium
Liquid；It is placed on high-pressure sterilizing pot to sterilize, 115 DEG C, 20min, for use；Take 34 parts by weight sterile waters and the above-mentioned culture medium of 1 parts by weight
Mixing, final culture medium Brix 2-3, pH 6.0.
(2) sample dilution and strain separating: by chocolate syrup (Brix60) (chocolate syrup flavor beverage, 2L/ bottles
(fresh and alive fruit juice Co., Ltd), Brix 59-61) 10 times of gradient dilutions of swell sample, take the sample 1ml of different dilutions with
Culture medium mixing inverted plate after cooling, while taking plate made of the sample scribing line culture medium of 200 microlitres of different gradients；Plate
It is placed in 35 DEG C of incubator cultures 3 days.
Comparative example 2 the difference from example 2 is that: culture medium Brix is not adjusted to 60.
To improvement and non-improved culture medium progress compliance test result: shown in the similar Fig. 1 and Fig. 2 of result, improved culture medium can be big
Amount screens the microorganism for causing swell, rather than improved culture medium screening has no that microorganism grows, and illustrates the improvement to culture medium
That is Brix adjustment is the key point of swell microorganism separation.
From reference Fig. 1-Fig. 2 and table 3 it is found that culture medium Brix made from the embodiment of the present invention 2 is adjusted to 60, osmotic pressure
It improves, which remains to grow, and can screen the preservative at pH4.0 and pH6.0 with fungistatic effect；And it is right
Ratio 2 has no that microorganism obviously grows.On the basis of improved culture medium, culture medium Brix is adjusted to 60, pH acid and neutral
Under the conditions of the poly- bad ammonia salt hydrochlorate of 0.01 parts by weight there is apparent fungistatic effect, sodium benzoate and potassium sorbate are in acid condition
The effect for inhibiting swell microorganism can be played.
In order to examine the actual effect for inhibiting swell, by 2.0 × 105Cfu/ml swell microorganism is linked into pH6.0's
In chocolate syrup product, addition or do not add 0.1 part of sodium benzoate, 35 DEG C insulating box culture 1 month, the results are shown in Table 4, swell
Microorganism 100% causes the chocolate syrup swell for not adding preservative sodium benzoate, and adds the chocolate of sodium benzoate
Slurry has no swell.
Influence whether adding sodium benzoate in 4 chocolate syrup of table to swell
| Project | Access strain viable count/cfu/ml | Swell quantity/ |
| 0.1 parts by weight sodium | 2.0×105 | 0/50 |
| Do not add sodium benzoate | 2.0×105 | 50/50 |
Therefore, the invention belongs to high sugared high magnification numbe beverage production technology fields, by conventional microbiological medium base
Upper progress Brix and pH is adjusted, and swell microorganism can be clearly separated.By adjusting fluid nutrient medium pH and Brix, imitates and produce
Product physicochemical environment can efficiently instruct the choosing of high sugar product preservative whether access certain amount swell microorganism observes its growth
It selects.The present invention can quickly screen objective microbe, and quickly can select most suitable preservative according to products characteristics.
To sum up, the microorganism separation for causing high sugar product swell of the invention and bacteriostasis method can be screened quickly and cause swell
Objective microbe, and obtain effective food preservative, it is ingenious in design to prevent the generation of swell, it is simple to operate, at
This is low, is suitable for large-scale promotion application.
It can be seen that the purpose of the present invention completely and is effectively achieved.Function and structural principle of the invention
It is shown and is illustrated in embodiment, under without departing substantially from the principle, embodiment can make any modification.So this hair
Bright includes all variant embodiments based on claim spirit and scope of the claims.
Claims (7)
1. a kind of microorganism separation for causing high sugar product swell and bacteriostasis method, which comprises the following steps:
(1) pol of microbiological culture media is adjusted to high pol 20Brix~60Brix, by the pH of the microbiological culture media
It adjusts to 4.0~6.0, obtains the microbiological culture media of improvement；
(2) by the microbiological culture media mixing inverted plate of the swell sample of high sugar product and the improvement, or will be described
The swell sample of high sugar product is crossed on the plate of the microbiological culture media of the improvement, and microculture is carried out；
(3) single colonie for selecting different shape carries out microorganism fungus kind identification, microorganism fungus kind qualification result is obtained, according to described
Microorganism fungus kind qualification result finds the suspicious microorganism for causing swell, and the suspicious microorganism is accessed the high sugar product
Non- swell sample in cultivated, observe the high sugar product non-swell sample whether can swell, if so, described can
Doubting microorganism is swell microorganism；
(4) different preservatives is separately added into the microbiological culture media of the fresh improvement at more parts and be inoculated with described swollen
Tank microorganism is cultivated and observes the swell microorganism whether there is or not growth, and selection inhibits the described anti-of the swell microorganism growth
Rotten agent.
2. causing microorganism separation and the bacteriostasis method of high sugar product swell as described in claim 1, which is characterized in that described
Step (1) specifically includes the following steps:
(11) take the solid component mixture of microbiological culture media described in 1 parts by weight, be added 7 parts by weight~21 parts by weight water with
And the HCl solution of 0 parts by weight~0.2 parts by weight 1mol/L, it is uniformly dissolved, obtains liquid medium；
(12) 1 parts by weight fructose syrup is taken, 0.04 other parts by weight~0.4 parts by weight water is added, is uniformly dissolved, obtains sugar
Liquid；
(13) liquid medium and the liquid glucose are individually positioned in high-pressure sterilizing pot 115 DEG C~121 DEG C, 15min~
20min sterilizing cools the mass ratio of rear 1:15~34 and mixes the liquid medium and the liquid glucose.
3. causing microorganism separation and the bacteriostasis method of high sugar product swell as described in claim 1, which is characterized in that in institute
It states in step (1), the microbiological culture media is plate count agar culture medium or rose bengal medium.
4. causing microorganism separation and the bacteriostasis method of high sugar product swell as described in claim 1, which is characterized in that described
Step (2) specifically includes the following steps:
(21) 10 times of gradient dilutions of the swell sample of the high sugar product are obtained into dilution；
(22) take microbiological culture media mixing inverted plate of the dilution described in 1ml with the improvement, or by 100 microlitres~
200 microlitres of dilution scribing line are on the plate of the microbiological culture media of the improvement；
(23) 35 DEG C are cultivated 1 day~3 days.
5. causing microorganism separation and the bacteriostasis method of high sugar product swell as described in claim 1, which is characterized in that in institute
It states in step (4), the pH of the microbiological culture media of the fresh improvement is 4 or 6.
6. causing microorganism separation and the bacteriostasis method of high sugar product swell as described in claim 1, which is characterized in that in institute
It states in step (4), the additive amount of the preservative is 0.01 parts by weight~0.1 parts by weight.
7. causing microorganism separation and the bacteriostasis method of high sugar product swell as described in claim 1, which is characterized in that in institute
It states in step (4), the preservative is selected from poly- two relied in ammonia salt hydrochlorate, nisin, potassium sorbate and sodium benzoate
Kind or more.
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| JP2010119364A (en) * | 2008-11-21 | 2010-06-03 | Koiwai Nyugyo Kk | Method for quickly detecting and measuring microorganism |
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Address after: 215321 No. 10 Jijin Road, Zhangpu Town, Kunshan City, Suzhou City, Jiangsu Province Applicant after: Fresh fruit juice Co.,Ltd. Address before: 215300 No. 10 Jijin Road, Zhangpu Town, Kunshan City, Suzhou City, Jiangsu Province Applicant before: SUNJUICE Co.,Ltd. Address after: 215321 No. 10 Jijin Road, Zhangpu Town, Kunshan City, Suzhou City, Jiangsu Province Applicant after: Suzhou fresh drink Co.,Ltd. Address before: 215321 No. 10 Jijin Road, Zhangpu Town, Kunshan City, Suzhou City, Jiangsu Province Applicant before: Fresh fruit juice Co.,Ltd. |
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